Wheat cultivar wb9518

ABSTRACT

The invention relates to the wheat cultivar designated WB9518. Provided by the invention are the seeds, plants and derivatives of the wheat cultivar WB9518. Also provided by the invention are tissue cultures of the wheat cultivar WB9518 and the plants regenerated therefrom. Still further provided by the invention are methods for producing wheat plants by crossing the wheat cultivar WB9518 with itself or another wheat cultivar and plants produced by such methods.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates generally to the field of wheat breeding.In particular, the invention relates to the new and distinctive wheatcultivar WB9518.

2. Description of Related Art

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possess the traits to meetthe program goals. The goal is to combine in a single variety animproved combination of desirable traits from the parental germplasm.These important traits may include higher seed yield, resistance todiseases and insects, better stems and roots, tolerance to drought andheat, better agronomic quality, resistance to herbicides, andimprovements in compositional traits.

Wheat may be classified into six different market classes. Five ofthese, including common wheat, hard red winter, hard red spring, softred winter, and white, belong to the species Triticum aestivum L., andthe sixth, durum, belongs to the species Triticum turgidum L. Wheat maybe used to produce a variety of products, including, but not limited to,grain, flour, baked goods, cereals, crackers, pasta, beverages,livestock feed, biofuel, straw, construction materials, and starches.The hard wheat classes are milled into flour used for breads, while thesoft wheat classes are milled into flour used for pastries and crackers.Wheat starch is used in the food and paper industries as laundrystarches, among other products.

SUMMARY OF THE INVENTION

One aspect of the present invention relates to seed of the wheatcultivar WB9518. The invention also relates to plants produced bygrowing the seed of the wheat cultivar WB9518, as well as thederivatives of such plants. Further provided are plant parts, includingcells, plant protoplasts, plant cells of a tissue culture from whichwheat plants can be regenerated, plant calli, plant clumps, and plantcells that are intact in plants or parts of plants, such as leaves,stems, roots, root tips, anthers, pistils, seed, grain, pericarp,embryo, pollen, ovules, cotyledon, hypocotyl, spike, floret, awn, lemma,shoot, tissue, petiole, cells, and meristematic cells, and the like.

In a further aspect, the invention provides a composition comprising aseed of wheat cultivar WB9518 comprised in plant seed growth media. Incertain embodiments, the plant seed growth media is a soil or syntheticcultivation medium. In specific embodiments, the growth medium may becomprised in a container or may, for example, be soil in a field. Plantseed growth media are well known to those of skill in the art andinclude, but are in no way limited to, soil or synthetic cultivationmedium. Advantageously, plant seed growth media can provide adequatephysical support for seeds and can retain moisture and/or nutritionalcomponents. Examples of characteristics for soils that may be desirablein certain embodiments can be found, for instance, in U.S. Pat. Nos.3,932,166 and 4,707,176. Synthetic plant cultivation media are also wellknown in the art and may, in certain embodiments, comprise polymers orhydrogels. Examples of such compositions are described, for example, inU.S. Pat. No. 4,241,537.

Another aspect of the invention relates to a tissue culture ofregenerable cells of the wheat cultivar WB9518, as well as plantsregenerated therefrom, wherein the regenerated wheat plant is capable ofexpressing all the physiological and morphological characteristics of aplant grown from the wheat seed designated WB9518.

Yet another aspect of the current invention is a wheat plant comprisinga single locus conversion of the wheat cultivar WB9518, wherein thewheat plant is otherwise capable of expressing all the physiological andmorphological characteristics of the wheat cultivar WB9518. Inparticular embodiments of the invention, the single locus conversion maycomprise a transgenic gene which has been introduced by genetictransformation into the wheat cultivar WB9518 or a progenitor thereof.In still other embodiments of the invention, the single locus conversionmay comprise a dominant or recessive allele. The locus conversion mayconfer potentially any trait upon the single locus converted plant,including, but not limited to, herbicide resistance, insect resistance,resistance to bacterial, fungal, or viral disease, male fertility orsterility, and improved nutritional quality.

Still yet another aspect of the invention relates to a first generation(F₁) hybrid wheat seed produced by crossing a plant of the wheatcultivar WB9518 to a second wheat plant. Also included in the inventionare the F₁ hybrid wheat plants grown from the hybrid seed produced bycrossing the wheat cultivar WB9518 to a second wheat plant. Stillfurther included in the invention are the seeds of an F₁ hybrid plantproduced with the wheat cultivar WB9518 as one parent, the secondgeneration (F₂) hybrid wheat plant grown from the seed of the F₁ hybridplant, and the seeds of the F₂ hybrid plant.

Still yet another aspect of the invention is a method of producing wheatseeds comprising crossing a plant of the wheat cultivar WB9518 to anysecond wheat plant, including itself or another plant of the cultivarWB9518. In particular embodiments of the invention, the method ofcrossing comprises the steps of: (a) planting seeds of the wheatcultivar WB9518; (b) cultivating wheat plants resulting from said seedsuntil said plants bear flowers; (c) allowing fertilization of theflowers of said plants; and (d) harvesting seeds produced from saidplants.

Still yet another aspect of the invention is a method of producinghybrid wheat seeds comprising crossing the wheat cultivar WB9518 to asecond, distinct wheat plant that is nonisogenic to the wheat cultivarWB9518. In particular embodiments of the invention, the crossingcomprises the steps of: (a) planting seeds of wheat cultivar WB9518 anda second, distinct wheat plant, (b) cultivating the wheat plants grownfrom the seeds until the plants bear flowers; (c) cross pollinating aflower on one of the two plants with the pollen of the other plant, and(d) harvesting the seeds resulting from the cross pollinating.

Still yet another aspect of the invention is a method for developing awheat plant in a wheat breeding program comprising: (a) obtaining awheat plant, or its parts, of the cultivar WB9518; and (b) employingsaid plant or parts as a source of breeding material using plantbreeding techniques. In the method, the plant breeding techniques may beselected from the group consisting of recurrent selection, massselection, bulk selection, backcrossing, pedigree breeding, geneticmarker-assisted selection and genetic transformation. In certainembodiments of the invention, the wheat plant of cultivar WB9518 may beused as the male or female parent.

Still yet another aspect of the invention is a method of producing awheat plant derived from the wheat cultivar WB9518, the methodcomprising the steps of: (a) preparing a progeny plant derived fromwheat cultivar WB9518 by crossing a plant of the wheat cultivar WB9518with a second wheat plant; and (b) crossing the progeny plant withitself or a second plant to produce a progeny plant of a subsequentgeneration which is derived from a plant of the wheat cultivar WB9518.In one embodiment of the invention, the method further comprises: (c)crossing the progeny plant of a subsequent generation with itself or asecond plant; and (d) repeating steps (b) and (c) for, in someembodiments, at least 2, 3, 4 or more additional generations to producean inbred wheat plant derived from the wheat cultivar WB9518. Alsoprovided by the invention is a plant produced by this and the othermethods of the invention.

In another embodiment of the invention, the method of producing a wheatplant derived from the wheat cultivar WB9518 further comprises: (a)crossing the wheat cultivar WB9518-derived wheat plant with itself oranother wheat plant to yield additional wheat cultivar WB9518-derivedprogeny wheat seed; (b) growing the progeny wheat seed of step (a) underplant growth conditions to yield additional wheat cultivarWB9518-derived wheat plants; and (c) repeating the crossing and growingsteps of (a) and (b) to generate further wheat cultivar WB9518-derivedwheat plants. In specific embodiments, steps (a) and (b) may be repeatedat least 1, 2, 3, 4, or 5 or more times as desired. The invention stillfurther provides a wheat plant produced by this and the foregoingmethods.

DETAILED DESCRIPTION OF THE INVENTION

Reference now will be made in detail to the embodiments of theinvention, one or more examples of which are set forth below. Eachexample is provided by way of explanation of the invention, not alimitation of the invention. It will be apparent to those skilled in theart that various modifications and variations may be made in the presentinvention without departing from the scope or spirit of the invention.For instance, features illustrated or described as part of oneembodiment, can be used on another embodiment to yield a still furtherembodiment.

Thus, it is intended that the present invention covers suchmodifications and variations as come within the scope of the appendedclaims and their equivalents. Other objects, features, and aspects ofthe present invention are disclosed in or are obvious from the followingdetailed description. It is to be understood by one of ordinary skill inthe art that the present discussion is a description of exemplaryembodiments only, and is not intended as limiting the broader aspects ofthe present invention.

In an embodiment, the invention is directed to wheat cultivar WB9518,its seeds, plants, and hybrids. Wheat cultivar WB9518 is a hard red,spring-type common wheat bred for spring planting in the hard red springwheat growing regions of the United States. The primary usage of wheatcultivar WB9518 is for production of grain, but it can also be used forproduction of silage harvested in the soft dough stage.

Breeding History

Wheat cultivar WB9518 is a common wheat cultivar developed from an F1selection made in 2007 from the cross of“‘TA/Solano//Expresso/3/Solano/4/Expresso/5/Hank’. “Solano,” “Expresso,”and “Hank” are commercial wheat varieties developed by WestBred LLC. TAis a symbol for a male sterile gene. The F1 was self-pollinated and thepedigree system of breeding was then used to develop and select WB9518,which is an F4 level selection. Some of the criteria used for selectionin various generations include yield, test weight, stripe rustresistance, % protein and SDS sedimentation score.

Wheat cultivar WB9518 has shown uniformity and stability in appearanceand performance across several generations (F5-F9) and acrossenvironments where it has been tested. The variety has been increasedwith continued observation for uniformity of plant type as described inthe following variety description information.

In accordance with another aspect of the invention, there is provided awheat plant having the physiological and morphological characteristicsof WB9518 as presented in Table 1. Those of skill in the art willrecognize that these are typical values that may vary due to environmentand that other values that are substantially equivalent are within thescope of the invention.

TABLE 1 Phenotypic Description of Wheat cultivar WB9518 CHARACTERISTICVALUES/RATINGS 1. PLANT WB9518 Coleoptile anthocyanin Absent Juvenileplant growth Erect Plant color at boot stage Blue-Green Flag leaf atboot stage Orientation Recurved Twist Twisted Waxy Bloom Present Days toHeading (Julian Date) 190 Anther color Yellow Plant height (cm) 80.4 2.STEM Anthocyanin Absent Waxy bloom Present Internode Form Hollow Number4 Hairiness of last internode of Present rachis Peduncle Form ErectLength (cm) 31 Auricle: Anthocyanin Absent Hair Present 3. HEAD (atmaturity) Density Middense Shape Strap Curvature Inclined AwnednessAwned 4. GLUMES (at maturity) Color White Shoulder Oblique Shoulderwidth Medium Beak shape Acuminate Beak width Medium (=1 mm) Glume lengthMedium (7.5-8.5 mm) Glume width Medium (3-4 mm) Pubescence Absent 5.SEED Shape Elliptical Cheek Angular Brush Medium Brush collar NotCollared Crease width Nearly as wide as kernel Crease depth 20% or lessof kernel Color Red Texture Hard Seed weight (g/1000 kernels) 31.5 Germsize Midsize Phenol reaction Dark Brown 6. DISEASE REACTIONS Pucciniagraminis f. sp. tritici (Stem Not Tested rust) Puccinia recondita f. sp.tritici (Leaf Not Tested rust) Puccinia striiformis (Stripe rust)Resistant Stagonospora nodorum (Glume blotch) Not Tested Septoriatritici (Speckled leaf blotch) Not Tested Erysiphe graminis f. sp.tritici (Powdery Not Tested mildew) Fusarium spp (Scab) IntermediateRhizoctonia solani Not Tested (Rhizoctonia root rot) Soil borne mosaicvirus (SBMV) Not Tested 7. INSECT REACTIONS Hessian fly (Mayetioladestructor) Tolerant

In an embodiment, the invention provides a composition comprising a seedof WB9518 comprised in plant seed growth media. Advantageously, plantseed growth media can provide adequate physical support for seeds andcan retain moisture and/or nutritional components. In certainembodiments, the plant seed growth media is a soil or syntheticcultivation medium. Any plant seed growth media known in the art may beutilized in this embodiment and the invention is in no way limited tosoil or synthetic cultivation medium. Examples of characteristics forsoils that may be desirable in certain embodiments can be found, forinstance, in U.S. Pat. Nos. 3,932,166 and 4,707,176. Plant cultivationmedia are well known in the art and may, in certain embodiments,comprise polymers, hydrogels, or the like. Examples of such compositionsare described, for example, in U.S. Pat. No. 4,241,537. In specificembodiments, the growth medium may be comprised in a container or may,for example, be soil in a field.

In another embodiment, the invention is directed to methods forproducing a wheat plant by crossing a first parent wheat plant with asecond parent wheat plant, wherein the first or second wheat plant isthe wheat plant from the cultivar WB9518. In an embodiment, the firstand second parent wheat plants may be from the cultivar WB9518 (i.e.,self-pollination). Any methods using the cultivar WB9518 are part ofthis invention: selfing, backcrosses, hybrid breeding, and crosses topopulations. Any plants produced using cultivar WB9518 as a parent arewithin the scope of this invention. In certain embodiments, theinvention is also directed to cells that, upon growth anddifferentiation, produce a cultivar having essentially all of thephysiological and morphological characteristics of WB9518. The presentinvention additionally contemplates, in various embodiments, a wheatplant regenerated from a tissue culture of cultivar WB9518.

In some embodiments of the invention, the invention is directed to atransgenic variant of WB9518. A transgenic variant of WB9518 may containat least one transgene but could contain at least 1, 2, 3, 4, 5, 6, 7,8, 9, 10, or more transgenes. In another embodiment, a transgenicvariant of WB9518 may contain no more than 15, 14, 13, 12, 11, 10, 9, 8,7, 6, 5, 4, 3, or 2 transgenes. Another embodiment of the inventioninvolves a process for producing wheat cultivar WB9518 furthercomprising a desired trait, said process comprising introducing atransgene that confers a desired trait to a wheat plant of cultivarWB9518. Methods for producing transgenic plants have been developed andare well known in the art. As part of the invention, one of ordinaryskill in the art may utilize any method of producing transgenic plantswhich is currently known or yet to be developed.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available.

In certain embodiments, the desired trait may be one or more ofherbicide tolerance or resistance, insect resistance or tolerance,disease resistance or tolerance, resistance for bacterial, viral, orfungal disease, male fertility, male sterility, decreased phytate, ormodified fatty acid or carbohydrate metabolism. The specific transgenemay be any known in the art or listed herein, including, but not limitedto a polynucleotide conferring resistance to imidazolinone, dicamba,sulfonylurea, glyphosate, glufosinate, triazine, benzonitrile,cyclohexanedione, phenoxy proprionic acid, and L-phosphinothricin; apolynucleotide encoding a Bacillus thuringiensis polypeptide, apolynucleotide encoding phytase, FAD-2, FAD-3, galactinol synthase or araffinose synthetic enzyme, Fusarium, Septoria, or various viruses orbacteria.

Various genetic elements can be introduced into the plant genome usingtransformation. These elements include, but are not limited to genes,coding sequences, inducible, constitutive, and tissue-specificpromoters, enhancing sequences, and signal and targeting sequences.

In some embodiments, the invention comprises a WB9518 plant that hasbeen developed using both genetic engineering and traditional breedingtechniques. For example, a genetic trait may have been engineered intothe genome of a particular wheat plant may then be moved into the genomeof a WB9518 plant using traditional breeding techniques that are wellknown in the plant breeding arts. Likewise, a genetic trait that hasbeen engineered into the genome of a WB9518 wheat plant may then bemoved into the genome of another variety using traditional breedingtechniques that are well known in the plant breeding arts. Abackcrossing approach is commonly used to move a transgene or transgenesfrom a transformed wheat variety into an already developed wheatvariety, and the resulting backcross conversion plant would thencomprise the transgene(s).

Plant transformation involves the construction of an expression vectorthat will function in plant cells. Such a vector may comprise DNAcomprising a gene under control of or operatively linked to a regulatoryelement (for example, a promoter). The expression vector may contain oneor more such operably linked gene/regulatory element combinations. Thevector(s) may be in the form of a plasmid and can be used alone or incombination with other plasmids to provide transformed wheat plants,using transformation methods as described below to incorporatetransgenes into the genetic material of the wheat plant(s).

Expression Vectors for Wheat Transformation: Marker Genes

Expression vectors may include at least one genetic marker operablylinked to a regulatory element that allows transformed cells containingthe marker to be either recovered by negative selection, i.e.,inhibiting growth of cells that do not contain the selectable markergene, or by positive selection, i.e., screening for the product encodedby the genetic marker. Many commonly used selectable marker genes forplant transformation are well known in the transformation arts, and mayinclude, for example, genes that code for enzymes that metabolicallydetoxify a selective chemical agent, which may be an antibiotic or anherbicide, or genes that encode an altered target which is insensitiveto the inhibitor. Positive selection methods are also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene, which when under thecontrol of plant regulatory signals, confers resistance to kanamycin.Another commonly used selectable marker gene is the hygromycinphosphotransferase gene, which confers resistance to the antibiotichygromycin.

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, and aminoglycoside-3′-adenyltransferase, the bleomycin resistance determinant. Other selectablemarker genes confer tolerance or resistance to herbicides such asglyphosate, glufosinate, or bromoxynil, or the like.

Other selectable marker genes for plant transformation that are not ofbacterial origin may include, for example, mouse dihydrofolatereductase, plant 5-enolpyruvylshikimate-3-phosphate synthase, and plantacetolactate synthase.

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells, rather than directgenetic selection of transformed cells, for resistance to a toxicsubstance such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase, and chloramphenicol acetyltransferase. Invivo methods for visualizing GUS activity that do not requiredestruction of plant tissue are also available. More recently, a geneencoding Green Fluorescent Protein (GFP) has been utilized as a markerfor gene expression in prokaryotic and eukaryotic cells. GFP and mutantsof GFP may also be used as screenable markers.

Expression Vectors for Wheat Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter. Manytypes of promoters are well known in the transformation arts, as areother regulatory elements that can be used alone or in combination withpromoters.

As used herein, “promoter” includes reference to a region of DNAupstream of the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters may be referred to as “tissue-preferred.”Promoters that initiate transcription only in certain tissue arereferred to as “tissue-specific.” A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter under environmental control. Examples of environmentalconditions that may affect transcription by inducible promoters includeanaerobic conditions or the presence of light. Tissue-specific,tissue-preferred, cell type specific, and inducible promoters constitutethe class of “non-constitutive” promoters. A “constitutive” promoter isa promoter that is active under most environmental conditions.

A. Inducible Promoters—An inducible promoter is operably linked to agene for expression in wheat. Optionally, the inducible promoter may beoperably linked to a nucleotide sequence encoding a signal sequence thatis operably linked to a gene for expression in wheat. With an induciblepromoter, the rate of transcription increases in response to an inducingagent.

Any inducible promoter may be used in the present invention. Exemplaryinducible promoters include, but are not limited to, those from the ACEIsystem, which respond to copper, and the In2 gene from maize, whichresponds to benzene-sulfonamide herbicide safeners. In an embodiment,the inducible promoter may be a promoter that responds to an inducingagent to which plants do not normally respond. An exemplary induciblepromoter may be an inducible promoter from a steroid hormone gene, thetranscriptional activity of which is induced by a glucocorticosteroidhormone.

B. Constitutive Promoters—A constitutive promoter is operably linked toa gene for expression in wheat, or is operably linked to a nucleotidesequence encoding a signal sequence that is operably linked to a genefor expression in wheat.

Many different constitutive promoters can be utilized in the presentinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses, such as the 35S promoter from CaMVand the promoters from such genes as rice actin; ubiquitin; pEMU; MAS,and maize H3 histone. The ALS promoter, Xba1/Nco1 fragment 5′ to theBrassica napus ALS3 structural gene (or a nucleotide sequence similarityto said Xba1/Nco1 fragment), represents a particularly usefulconstitutive promoter.

C. Tissue-specific or Tissue-preferred Promoters—A tissue-specificpromoter is operably linked to a gene for expression in wheat. Thetissue-specific promoter may be operably linked to a nucleotide sequenceencoding a signal sequence that is operably linked to a gene forexpression in wheat. Plants transformed with a gene of interest operablylinked to a tissue-specific promoter may produce the protein product ofthe transgene exclusively, or preferentially, in a specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in thepresent invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferred promoter,such as that from the phaseolin gene; a leaf-specific and light-inducedpromoter, such as that from cab or rubisco; an anther-specific promoter,such as that from LAT52; a pollen-specific promoter, such as that fromZml 3; or a microspore-preferred promoter, such as that from apg.

Signal Sequences for Targeting Proteins to Subcellular Compartments

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall, ormitochondrion, or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized. The presence of asignal sequence directs a polypeptide to either an intracellularorganelle or subcellular compartment or for secretion to the apoplast.Many signal sequences are known in the art.

Foreign Protein Genes and Agronomic Genes

Using transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants that maybe harvested in a conventional manner. A foreign protein can then beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods.

According to an embodiment of the invention, the transgenic plantprovided for commercial production of foreign protein is, or is derivedfrom a WB9518 wheat plant. In another embodiment, the biomass ofinterest is or is derived from a WB9518 seed. For the relatively smallnumber of transgenic plants that show higher levels of expression, agenetic map can be generated, primarily via conventional restrictionfragment length polymorphism (RFLP), polymerase chain reaction (PCR),and simple sequence repeat (SSR) analysis, which identify theapproximate chromosomal location of the integrated DNA molecule. Mapinformation concerning chromosomal location is useful for proprietaryprotection of a subject transgenic plant. If unauthorized propagation isundertaken and crosses made with other germplasm, the map of theintegration region can be compared to similar maps for suspect plants,to determine if the latter have a common parentage with the subjectplant. Map comparisons would involve hybridizations, RFLP, PCR, SSR, andsequencing, all of which are conventional techniques well known in theart.

In certain embodiments, the invention comprises transformed WB9518plants that express particular agronomic genes or phenotypes ofagronomic interest. Exemplary genes implicated in this regard include,but are not limited to, those categorized below:

1. Genes that Confer Tolerance or Resistance to Pests or Disease andthat Encode:

A. Plant disease tolerance or resistance genes. Plant defenses are oftenactivated by specific interaction between the product of a diseasetolerance or resistance gene (R) in the plant and the product of acorresponding avirulence (Avr) gene in the pathogen. A plant variety canbe transformed with one or more cloned resistance genes to engineerplants that are resistant to specific pathogen strains.

B. A gene conferring resistance to a pest, such as nematodes.

C. A Bacillus thuringiensis protein, a derivative thereof, or asynthetic polypeptide modeled thereon. Moreover, DNA molecules encoding8-endotoxin genes can be purchased from American Type CultureCollection, Manassas, Va., for example, under ATCC Accession Nos. 40098,67136, 31995 and 31998.

D. A lectin. The nucleotide sequence of several Clivia miniatamannose-binding lectin genes are known in the art.

E. A vitamin-binding protein such as avidin or avidin homologues.

F. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. For example, the nucleotide sequences of ricecysteine proteinase inhibitor, cDNA encoding tobacco proteinaseinhibitor I, and Streptomyces nitrosporeus α-amylase inhibitor are knownin the art.

G. An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. For example, the baculovirus expressionof cloned juvenile hormone esterase, an inactivator of juvenile hormone,is known in the art.

H. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, it is knownthat expression cloning yields DNA coding for insect diuretic hormonereceptor and an allostatin can be identified in Diploptera puntata.Genes encoding insect-specific, paralytic neurotoxins are also known inthe art.

I. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, heterologous expression in plants of a gene coding for ascorpion insectotoxic peptide is known in the art.

J. An enzyme responsible for a hyper-accumulation of a monoterpene, asesquiterpene, a steroid, a hydroxamic acid, a phenylpropanoidderivative, or another non-protein molecule with insecticidal activity.

K. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule. Forexample, such enzymes include, but are not limited to, a glycolyticenzyme, a proteolytic enzyme, a lipolytic enzyme, a nuclease, a cyclase,a callase, a transaminase, an esterase, a hydrolase, a phosphatase, akinase, a phosphorylase, a polymerase, an elastase, a chitinase, and aglucanase, whether natural or synthetic. DNA molecules that containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. The nucleotide sequences of a cDNAencoding tobacco hookworm chitinase and parsley ubi4-2 polyubiquitingene are also known in the art.

L. A molecule that stimulates signal transduction. For example, thenucleotide sequences for mung bean calmodulin cDNA clones and a maizecalmodulin cDNA clone are known in the art.

M. A hydrophobic moment peptide. For example, peptide derivatives ofTachyplesin, which inhibit fungal plant pathogens, or syntheticantimicrobial peptides that confer disease resistance.

N. A membrane permease, a channel former, or a channel blocker. Forexample, heterologous expression of a cecropin-13 lytic peptide analogto render transgenic tobacco plants resistant to Pseudomonassolanacearum is known in the art.

O. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmentaffected by the virus from which the coat protein gene is derived, aswell as by related viruses. Coat protein-mediated resistance has beenconferred upon transformed plants against alfalfa mosaic virus, cucumbermosaic virus, tobacco streak virus, potato virus X, potato virus Y,tobacco etch virus, tobacco rattle virus, and tobacco mosaic virus.

P. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Forexample, enzymatic inactivation in transgenic tobacco via production ofsingle-chain antibody fragments is known in the art.

Q. A virus-specific antibody. For example, it is known in the art thattransgenic plants expressing recombinant antibody genes are protectedfrom virus attack.

R. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo-α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. The cloning and characterizationof a gene that encodes a bean endopolygalacturonase-inhibiting proteinis known in the art.

S. A developmental-arrestive protein produced in nature by a plant. Forexample, it has been shown that transgenic plants expressing the barleyribosome-inactivating gene have an increased resistance to fungaldisease.

T. Genes expressing proteins with antifungal action. Fusarium headblight along with deoxynivalenol both produced by the pathogen Fusariumgraminearum (Schwabe) have caused devastating losses in wheatproduction. Genes expressing proteins with antifungal action can be usedas transgenes to prevent Fusarium head blight. Various classes ofproteins have been identified. Examples include endochitinases,exochitinases, glucanases, thionins, thaumatin-like proteins, osmotins,ribosome-inactivating proteins, flavonoids, and lactoferricin. Duringinfection with Fusarium graminearum, deoxynivalenol is produced. Thereis evidence that production of deoxynivalenol increases the virulence ofthe disease. Genes with properties for detoxification of deoxynivalenolhave been engineered for use in wheat. A synthetic peptide that competeswith deoxynivalenol has been identified. Changing the ribosomes of thehost so that they have reduced affinity for deoxynivalenol has also beenused to reduce the virulence of Fusarium graminearum. Genes used to helpreduce Fusarium head blight include, but are not limited to, Tri101(Fusarium), PDR5 (yeast), tlp-1 (oat), tlp-2 (oat), leaf tlp-1 (wheat),tlp (rice), tlp-4 (oat), endochitinase, exochitinase, glucanase(Fusarium), permatin (oat), seed hordothionin (barley), alpha-thionin(wheat), acid glucanase (alfalfa), chitinase (barley and rice), classbeta II-1,3-glucanase (barley), PR5/tlp (Arabidopsis), zeamatin (maize),type 1 RIP (barley), NPR1 (Arabidopsis), lactoferrin (mammal),oxalylCoA-decarboxylase (bacterium), IAP (baculovirus), ced-9 (C.elegans), and glucanase (rice and barley).

U. A gene, for example, the H9, H10, and H21 genes, conferringresistance to a pest, such as Hessian fly, stem soft fly, cereal leafbeetle, and/or green bug.

V. A gene conferring resistance to diseases such as wheat rusts,Septoria tritici, Septoria nodorum, powdery mildew, Helminthosporiumdiseases, smuts, bunts, Fusarium diseases, bacterial diseases, and viraldiseases.

W. Genes involved in the Systemic Acquired Resistance (SAR) responseand/or the pathogenesis-related genes.

X. Antifungal genes.

Y Detoxification genes, such as for fumonisin, beauvericin,moniliformin, and zearalenone and their structurally relatedderivatives.

Z. Cystatin and cysteine proteinase inhibitors.

AA. Defensin genes.

BB. Genes conferring resistance to nematodes.

2. Genes that Confer Tolerance or Resistance to an Herbicide:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme.

B. Glyphosate (resistance conferred by mutant5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and aroA genes) andother phosphono compounds, such as glufosinate (phosphinothricin acetyltransferase (PAT) and Streptomyces hygroscopicus PAT bar genes), andpyridinoxy or phenoxy proprionic acids and cyclohexones (ACCaseinhibitor-encoding genes). For example, the nucleotide sequence of aform of EPSP which can confer glyphosate resistance is known in the art.A DNA molecule encoding a mutant aroA gene can be obtained under ATCCaccession number 39256, and the nucleotide sequence of the mutant geneis known. Nucleotide sequences of glutamine synthetase genes that confertolerance or resistance to herbicides such as L-phosphinothricin arealso known in the art. The nucleotide sequence of a PAT gene is known inthe art, as is the production of transgenic plants that express chimericbar genes coding for PAT activity. Exemplary genes conferring resistanceto phenoxy proprionic acids and cyclohexones, such as sethoxydim andhaloxyfop are the Accl-S1, Accl-S2, and Accl-S3 genes.

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+genes) or a benzonitrile (nitrilase gene). Nucleotide sequencesfor nitrilase genes are disclosed and DNA molecules containing thesegenes are available under ATCC Accession Nos. 53435, 67441, and 67442.Cloning and expression of DNA coding for a glutathione S-transferase isdescribed in the art.

D. Acetohydroxy acid synthase. This enzyme has been found to make plantsthat express this enzyme tolerant or resistant to multiple types ofherbicides and has been introduced into a variety of plants. Other genesthat confer tolerance or resistance to herbicides include a geneencoding a chimeric protein of rat cytochrome P4507A1 and yeastNADPH-cytochrome P450 oxidoreductase, genes for glutathione reductaseand superoxide dismutase, and genes for various phosphotransferases.

E. Protoporphyrinogen oxidase (protox). Protox is necessary for theproduction of chlorophyll, which is necessary for survival in allplants. The protox enzyme serves as the target for a variety ofherbicidal compounds. These herbicides also inhibit growth of differentspecies of plants present, causing their total destruction. Thedevelopment of plants containing altered protox activity that aretolerant or resistant to these herbicides is described in the art.

3. Genes that Confer or Contribute to a Value-Added Trait, Such as:

A. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant.

B. Decreased phytate content. This can be accomplished by: (1)Introduction of a phytase-encoding gene that enhances breakdown ofphytate, adding more free phosphate to the transformed plant; or (2)Up-regulation of a gene that reduces phytate content. For example, thenucleotide sequence of an Aspergillus niger phytase gene has beendescribed in the art.

C. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch, or, a gene altering thioredoxin such as NTRand/or TRX and/or a gamma zein knockout or mutant, such as cs27, TUSC27,or en27. For example, the nucleotide sequences of Streptococcus mutansfructosyltransferase gene, Bacillus subtilis levansucrase gene, andtomato invertase genes are known in the art. Transgenic plants can beproduced that express Bacillus licheniformis alpha-amylase, thatsite-direct mutagenesis of barley alpha-amylase gene, or confer maizeendosperm starch branching enzyme II or improved digestibility and/orstarch extraction through modification of UDP-D-xylose 4-epimerase.Methods of producing high oil seed by modification of starch levels(AGP) are also known. The fatty acid modification genes mentioned abovemay also be used to affect starch content and/or composition through theinterrelationship of the starch and oil pathways.

D. Elevated oleic acid via FAD-2 gene modification and/or decreasedlinolenic acid via FAD-3 gene modification.

E. Altering conjugated linolenic or linoleic acid content, or LEC1, AGP,Dekl, Superall, milps, various Ipa genes such as Ipa1, Ipa3, hpt, orhggt.

F. Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, manipulation of antioxidantlevels through alteration of a phytl prenyl transferase (ppt) is known,as is manipulation of antioxidant levels through alteration of ahomogentisate geranyl geranyl transferase (hggt).

G. The content of high-molecular weight gluten subunits (HMS-GS).Genomic clones have been isolated for different subunits. For example,genomic clones have transformed wheat with genes that encode a modifiedHMW-GS.

H. Increased protein metabolism, zinc and iron content, for example, byregulating the NAC gene, increasing protein metabolism by regulating theGpc-B 1 gene, or regulating glutenin and gliadin genes.

I. Altered essential seed amino acids. Methods of increasingaccumulation of essential amino acids in seeds, binary methods ofincreasing accumulation of essential amino acids in seeds, alteration ofamino acid compositions in seeds, methods for altering amino acidcontent of proteins, alteration of amino acid compositions in seeds, andproteins with enhanced levels of essential amino acids all are known inthe art. Other examples may include high methionine, high threonine,plant amino acid biosynthetic enzymes, increased lysine and threonine,plant tryptophan synthase beta subunit, methionine metabolic enzymes,high sulfur, increased methionine, plant amino acid biosyntheticenzymes, engineered seed protein having higher percentage of essentialamino acids, increased lysine, increasing sulfur amino acid content,synthetic storage proteins with defined structure containingprogrammable levels of essential amino acids for improvement of thenutritional value of plants, increased threonine, increased lysine, CesA: cellulose synthase, hemicellulose, UDPGdH, and RGP.

4. Genes that Control Male Sterility

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility. In addition to these methods, asystem of nuclear male sterility that includes: identifying a gene whichis critical to male fertility; silencing this native gene which iscritical to male fertility; removing the native promoter from theessential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on”, resulting in the male fertility gene notbeing transcribed, is known. Fertility is restored by inducing, orturning “on”, the promoter, which in turn allows the gene that confersmale fertility to be transcribed.

A. Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN—Ac-PPT.

B. Introduction of various stamen-specific promoters.

C. Introduction of the barnase and the barstar genes.

5. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.Other systems that may be used include the Gin recombinase of phage Mu,the Pin recombinase of E. coli, and the R/RS system of the pSRi plasmid.

6. Genes that Affect Abiotic Stress Resistance.

A. Genes that affect abiotic stress resistance (including but notlimited to flowering, seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress. For example,water use efficiency can be altered through alteration of malate. Inaddition, various genes, including CBF genes and transcription factors,can be effective in mitigating the negative effects of freezing, highsalinity, and drought on plants, as well as conferring other positiveeffects on plant phenotype. Abscisic acid can be altered in plants,resulting in improved plant phenotype, such as increased yield and/orincreased tolerance to abiotic stress. Cytokinin expression can bemodified resulting in plants with increased stress tolerance, such asdrought tolerance, and/or increased yield. Nitrogen utilization can beenhanced and/or nitrogen responsiveness can be altered. Ethylene can bealtered. Plant transcription factors or transcriptional regulators ofabiotic stress can also be altered.

B. Improved tolerance to water stress from drought or high salt watercondition. The HVA1 protein belongs to the group 3 LEA proteins thatinclude other members such as wheat pMA2005, cotton D-7, carrot Dc3, andrape pLEA76. These proteins are characterized by 11-mer tandem repeatsof amino acid domains which may form a probable amphophilicalpha-helical structure that presents a hydrophilic surface with ahydrophobic stripe. The barley HVA1 gene and the wheat pMA2005 gene arehighly similar at both the nucleotide level and predicted amino acidlevel. These two monocot genes are closely related to the cotton D-7gene and carrot Dc3 gene with which they share a similar structural geneorganization. There is, therefore, a correlation between LEA geneexpression or LEA protein accumulation with stress tolerance in a numberof plants. For example, in severely dehydrated wheat seedlings, theaccumulation of high levels of group 3 LEA proteins was correlated withtissue dehydration tolerance. Studies on several Indica varieties ofrice showed that the levels of group 2 LEA proteins (also known asdehydrins) and group 3 LEA proteins in roots were significantly higherin salt-tolerant varieties compared with sensitive varieties. The barleyHVA1 gene was transformed into wheat. Transformed wheat plants showedincreased tolerance to water stress.

C. Improved water stress tolerance through increased mannitol levels viathe bacterial mannitol-1-phosphate dehydrogenase gene. It is known toproduce a plant with a genetic basis for coping with water deficit byintroduction of the bacterial mannitol-1-phosphate dehydrogenase gene,mt1D, into tobacco cells via Agrobacterium-mediated transformation. Rootand leaf tissues from transgenic plants regenerated from thesetransformed tobacco cells contained up to 100 mM mannitol. Controlplants contained no detectable mannitol. To determine whether thetransgenic tobacco plants exhibited increased tolerance to waterdeficit, the growth of transgenic plants was compared to that ofuntransformed control plants in the presence of 250 mM NaCl. After 30days of exposure to 250 mM NaCl, transgenic plants had decreased weightloss and increased height relative to their untransformed counterparts.The authors concluded that the presence of mannitol in these transformedtobacco plants contributed to water deficit tolerance at the cellularlevel.

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants.

Methods for Wheat Transformation

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. In addition,expression vectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available.

A. Agrobacterium-Mediated Transformation. One method for introducing anexpression vector into plants is based on the natural transformationsystem of Agrobacterium. A. tumefaciens and A. rhizogenes are plantpathogenic soil bacteria that genetically transform plant cells. The Tiand Ri plasmids of A. tumefaciens and A. rhizogenes, respectively, carrygenes responsible for genetic transformation of the plant. Descriptionsof Agrobacterium vector systems and methods for Agrobacterium-mediatedgene transfer are well known in the art.

B. Direct Gene Transfer. Several methods of plant transformation,collectively referred to as direct gene transfer, have been developed asan alternative to Agrobacterium-mediated transformation. A generallyapplicable method of plant transformation is microprojectile-mediatedtransformation, wherein DNA is carried on the surface ofmicroprojectiles measuring 1 to 4 pm. The expression vector isintroduced into plant tissues with a biolistic device that acceleratesthe microprojectiles to speeds of 300 to 600 m/s, which is sufficient topenetrate plant cell walls and membranes.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Alternatively, liposome and spheroplast fusion have beenused to introduce expression vectors into plants. Direct uptake of DNAinto protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpolyL-ornithine has also been reported. Electroporation of protoplastsand whole cells and tissues has also been described. Followingtransformation of wheat target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods that are well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic variety. The transgenic variety could then becrossed, with another (non-transformed or transformed) variety, in orderto produce a new transgenic variety. Alternatively, a genetic traitwhich has been engineered into a particular wheat cultivar using theforegoing transformation techniques could be moved into another cultivarusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite variety into anelite variety, or from a variety containing a foreign gene in its genomeinto a variety or varieties which do not contain that gene. As usedherein, “crossing” can refer to a simple X by Y cross, or the process ofbackcrossing, depending on the context.

Genetic Marker Profile Through SSR and First Generation Progeny

In addition to phenotypic observations, a plant can also be identifiedby its genotype. The genotype of a plant can be characterized through agenetic marker profile, which can identify plants of the same variety ora related variety or be used to determine or validate a pedigree.Genetic marker profiles can be obtained by techniques such asRestriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asmicrosatellites, and Single Nucleotide Polymorphisms (SNPs).

Particular markers used for these purposes are not limited to anyparticular set of markers, but are envisioned to include any type ofmarker and marker profile that provides a means of distinguishingvarieties. One method of comparison is to use only homozygous loci forWB9518.

In addition to being used for identification of wheat cultivar WB9518and plant parts and plant cells of cultivar WB9518, the genetic profilemay be used to identify a wheat plant produced through the use of WB9518or to verify a pedigree for progeny plants produced through the use ofWB9518. The genetic marker profile is also useful in breeding anddeveloping backcross conversions.

In some embodiments, the present invention comprises a wheat plantcharacterized by molecular and physiological data obtained from therepresentative sample of WB9518, deposited with the American TypeCulture Collection (ATCC). Provided in further embodiments of theinvention is a wheat plant formed by the combination of the WB9518 plantor plant cell with another wheat plant or cell and comprising thehomozygous alleles of the variety.

Means of performing genetic marker profiles using SSR polymorphisms arewell known in the art. SSRs are genetic markers based on polymorphismsin repeated nucleotide sequences, such as microsatellites. A markersystem based on SSRs can be highly informative in linkage analysisrelative to other marker systems in that multiple alleles may bepresent. Another advantage of this type of marker is that, through useof flanking primers, detection of SSRs can be achieved, for example, bythe polymerase chain reaction (PCR), thereby eliminating the need forlabor-intensive Southern hybridization. PCR detection uses twooligonucleotide primers flanking the polymorphic segment of repetitiveDNA. Repeated cycles of heat denaturation of the DNA, followed byannealing of the primers to their complementary sequences at lowtemperatures, and extension of the annealed primers with DNA polymerase,comprise the major part of the methodology.

Following amplification, markers can be scored by electrophoresis of theamplification products. Scoring of marker genotype is based on the sizeof the amplified fragment, which may be measured by the number of basepairs of the fragment. While variation in the primer used or inlaboratory procedures can affect the reported fragment size, relativevalues should remain constant regardless of the specific primer orlaboratory used. When comparing varieties, all SSR profiles may beperformed in the same lab.

The SSR profile of wheat plant WB9518 can be used to identify plantscomprising WB9518 as a parent, since such plants will comprise the samehomozygous alleles as WB9518. Because the wheat variety is essentiallyhomozygous at all relevant loci, most loci should have only one type ofallele present. In contrast, a genetic marker profile of an F1 progenyshould be the sum of those parents, e.g., if one parent was homozygousfor allele x at a particular locus, and the other parent homozygous forallele y at that locus, then the F1 progeny will be xy (heterozygous) atthat locus. Subsequent generations of progeny produced by selection andbreeding are expected to be of genotype x (homozygous), y (homozygous),or xy (heterozygous) for that locus position. When the F1 plant isselfed or sibbed for successive filial generations, the locus should beeither x or y for that position.

In addition, plants and plant parts substantially benefiting from theuse of WB9518 in their development, such as WB9518 comprising abackcross conversion, transgene, or genetic sterility factor, may beidentified by having a molecular marker profile with a high percentidentity to WB9518. In an embodiment, such a percent identity might be95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to WB9518.

The SSR profile of WB9518 also can be used to identify essentiallyderived varieties and other progeny varieties developed from the use ofWB9518, as well as cells and other plant parts thereof. Progeny plantsand plant parts produced using WB9518 may be identified by having amolecular marker profile of at least 25%, 30%, 35%, 40%, 45%, 50%, 55%,60%, 65%, 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or99.5% genetic contribution fromWB9518, as measured by either percentidentity or percent similarity. Such progeny may be furthercharacterized as being within a pedigree distance of WB9518, such aswithin 1, 2, 3, 4 or 5 or fewer cross-pollinations to a wheat plantother than WB9518 or a plant that has WB9518 as a progenitor. Uniquemolecular profiles may be identified with other molecular tools such asSNPs and RFLPs.

While determining the SSR genetic marker profile of a plant as describedabove, several unique SSR profiles may also be identified that did notappear in either parent plant. Such unique SSR profiles may arise duringthe breeding process from recombination or mutation. A combination ofseveral unique alleles provides a means of identifying a plant variety,an F1 progeny produced from such variety, and further progeny producedfrom such variety.

Gene Conversion

When the term “wheat plant” is used in the context of the presentinvention, this also includes any gene conversions of that variety.Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the variety. For example, a varietymay be backcrossed 1, 2, 3, 4, 5, 6, 7, 8, 9 or more times to therecurrent parent. The parental wheat plant that contributes the gene forthe desired characteristic is termed the “nonrecurrent” or “donor”parent. This terminology refers to the fact that the nonrecurrent parentis used one time in the backcross protocol and therefore does not recur.The parental wheat plant to which the gene or genes from thenonrecurrent parent are transferred is known as the recurrent parent, asit is used for several rounds in the backcrossing protocol. In a typicalbackcross protocol, the original variety of interest (recurrent parent)is crossed to a second variety (nonrecurrent parent) that carries thesingle gene of interest to be transferred. The resulting progeny fromthis cross are then crossed again to the recurrent parent and theprocess is repeated until a wheat plant is obtained wherein essentiallyall of the desired morphological and physiological characteristics ofthe recurrent parent are recovered in the converted plant, in additionto the single transferred gene from the nonrecurrent parent.

The selection of a suitable recurrent parent contributes to a successfulbackcrossing procedure. The goal of a backcross protocol is to alter orsubstitute a single trait or characteristic in the original variety. Toaccomplish this, a single gene of the recurrent variety is modified orsubstituted with the desired gene from the nonrecurrent parent, whileretaining essentially all of the rest of the desired genetic, andtherefore the desired physiological and morphological, constitution ofthe original variety. The choice of the particular nonrecurrent parentwill depend on the purpose of the backcross. One of the major purposesis to add commercially desirable, agronomically important traits to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele may also be transferred. In this instance, it may benecessary to introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

Many single gene traits have been identified that are not regularlyselected for in the development of a new variety, but that can beimproved by backcrossing techniques. Single gene traits may or may notbe transgenic. Examples of these traits include, but are not limited to,male sterility, waxy starch, herbicide tolerance or resistance,resistance for bacterial, fungal, or viral disease, insect resistance ortolerance, male fertility, enhanced nutritional quality, industrialusage, yield stability and yield enhancement. These genes are generallyinherited through the nucleus.

Introduction of a New Trait or Locus into WB9518

Cultivar WB9518 represents a new base genetic variety into which a newlocus or trait may be introgressed. Direct transformation andbackcrossing represent two important methods that can be used toaccomplish such an introgression. The term backcross conversion andsingle locus conversion are used interchangeably to designate theproduct of a backcrossing program.

Backcross Conversions of WB9518

A backcross conversion of WB9518 occurs when DNA sequences areintroduced through backcrossing, with WB9518 utilized as the recurrentparent. Both naturally occurring and transgenic DNA sequences may beintroduced through backcrossing techniques. A backcross conversion mayproduce a plant with a trait or locus conversion in at least two or morebackcrosses, including at least 2 crosses, at least 3 crosses, at least4 crosses, at least 5 crosses, or additional crosses. Molecular markerassisted breeding or selection may be utilized to reduce the number ofbackcrosses necessary to achieve the backcross conversion. For example,a backcross conversion can be made in as few as two backcrosses.

The complexity of the backcross conversion method depends on the type oftrait being transferred (single genes or closely linked genes versusunlinked genes), the level of expression of the trait, the type ofinheritance (cytoplasmic or nuclear) and the types of parents includedin the cross. It is understood by those of ordinary skill in the artthat for single gene traits that are relatively easy to classify, thebackcross method is effective and relatively easy to manage. Desiredtraits that may be transferred through backcross conversion include, butare not limited to, sterility (nuclear and cytoplasmic), fertilityrestoration, nutritional enhancements, drought tolerance, nitrogenutilization, altered fatty acid profile, low phytate, industrialenhancements, disease resistance or tolerance (bacterial, fungal orviral), insect resistance or tolerance, and herbicide tolerance orresistance. In addition, an introgression site itself, such as an FRTsite, Lox site, or other site specific integration site, may be insertedby backcrossing and utilized for direct insertion of one or more genesof interest into a specific plant variety. In some embodiments of theinvention, the number of loci that may be backcrossed into WB9518 is atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more. A single locus may containseveral transgenes, such as a transgene for disease resistance that, inthe same expression vector, also contains a transgene for herbicidetolerance or resistance. The gene for herbicide tolerance or resistancemay be used as a selectable marker and/or as a phenotypic trait. Asingle locus conversion of site specific integration system allows forthe integration of multiple genes at the converted loci.

The backcross conversion may result from either the transfer of adominant allele or a recessive allele. Selection of progeny containingthe trait of interest is accomplished by direct selection for a traitassociated with a dominant allele. Transgenes transferred viabackcrossing typically function as a dominant single gene trait and arerelatively easy to classify. Selection of progeny for a trait that istransferred via a recessive allele requires growing and selling thefirst backcross generation to determine which plants carry the recessivealleles. Recessive traits may require additional progeny testing insuccessive backcross generations to determine the presence of the locusof interest. The last backcross generation is usually selfed to givepure breeding progeny for the gene(s) being transferred, although abackcross conversion with a stably introgressed trait may also bemaintained by further backcrossing to the recurrent parent withselection for the converted trait.

Along with selection for the trait of interest, progeny are selected forthe phenotype of the recurrent parent. The backcross is a form ofinbreeding, and the features of the recurrent parent are automaticallyrecovered after successive backcrosses. Some sources suggest from one tofour or more backcrosses, but as noted above, the number of backcrossesnecessary can be reduced with the use of molecular markers. Otherfactors, such as a genetically similar donor parent, may also reduce thenumber of backcrosses necessary. Backcrossing is easiest for simplyinherited, dominant and easily recognized traits.

One process for adding or modifying a trait or locus in wheat cultivarWB9518 comprises crossing WB9518 plants grown from WB9518 seed withplants of another wheat variety that comprise the desired trait orlocus, selecting F1 progeny plants that comprise the desired trait orlocus to produce selected F1 progeny plants, crossing the selectedprogeny plants with the WB9518 plants to produce backcross progenyplants, selecting for backcross progeny plants that have the desiredtrait or locus and the morphological characteristics of wheat cultivarWB9518 to produce selected backcross progeny plants, and backcrossing toWB9518 three or more times in succession to produce selected fourth orhigher backcross progeny plants that comprise said trait or locus. Themodified WB9518 may be further characterized as having essentially allof the physiological and morphological characteristics of wheat cultivarWB9518 listed in Table 1, as determined at the 5% significance levelwhen grown in the same environmental conditions and/or may becharacterized by percent similarity or identity to WB9518 as determinedby SSR markers. The above method may be utilized with fewer backcrossesin appropriate situations, such as when the donor parent is highlyrelated or markers are used in the selection step. Desired nucleic acidsthat may be used include those nucleic acids known in the art, some ofwhich are listed herein, that will affect traits through nucleic acidexpression or inhibition. Desired loci include the introgression of FRT,Lox, and other sites for site specific integration, which may alsoaffect a desired trait if a functional nucleic acid is inserted at theintegration site.

In addition, the above process and other similar processes describedherein may be used to produce first generation progeny wheat seed byadding a step at the end of the process that comprises crossing WB9518with the introgressed trait or locus with a different wheat plant andharvesting the resultant first generation progeny wheat seed.

A further embodiment of the invention is a back-cross conversion ofwheat cultivar WB9518. A backcross conversion occurs when DNA sequencesare introduced through traditional (non-transformation) breedingtechniques, such as backcrossing. DNA sequences, whether naturallyoccurring or transgenes, may be introduced using these traditionalbreeding techniques. Desired traits transferred through this processinclude, but are not limited to nutritional enhancements, industrialenhancements, disease resistance or tolerance, insect resistance ortolerance, herbicide tolerance or resistance, agronomic enhancements,grain quality enhancement, waxy starch, breeding enhancements, seedproduction enhancements, and male sterility. Descriptions of some of thecytoplasmic male sterility genes, nuclear male sterility genes, chemicalhybridizing agents, male fertility restoration genes, and methods ofusing the aforementioned are known. Examples of genes for other traitsinclude: Leaf rust resistance genes (Lr series such as Lr1, Lr10, Lr21,Lr22, Lr22a, Lr32, Lr37, Lr41, Lr42, and Lr43), Fusarium headblight-resistance genes (QFhs.ndsu-3B and QFhs.ndsu-2A), powdery mildewresistance genes (Pm21), common bunt resistance genes (Bt-10), and wheatstreak mosaic virus resistance gene (Wsm1), Russian wheat aphidresistance genes (Dn series such as Dn1, Dn2, Dn4, and Dn5), Black stemrust resistance genes (Sr38), Yellow rust resistance genes (Yr seriessuch as Yr 1, YrSD, Yrsu, Yr17, Yr15, and YrH52), aluminum tolerancegenes (Alt(BH)), dwarf genes (Rht), vernalization genes (Vrn), Hessianfly resistance genes (H9, H10, H21, and H29), grain color genes (R/r),glyphosate resistance genes (EPSPS), glufosinate genes (bar, pat) andwater stress tolerance genes (Hva 1 and mt1D). The trait of interest istransferred from the donor parent to the recurrent parent, which in thiscase is the wheat plant disclosed herein, WB9518. Single gene traits mayresult from either the transfer of a dominant allele or a recessiveallele. Selection of progeny containing the trait of interest is done bydirect selection for a trait associated with a dominant allele.Selection of progeny for a trait that is transferred via a recessiveallele requires growing and selfing the first backcross to determinewhich plants carry the recessive alleles. Recessive traits may requireadditional progeny testing in successive backcross generations todetermine the presence of the gene of interest.

Using WB9518 to Develop Other Wheat Varieties

Wheat varieties such as WB9518 are typically developed for use in seedand grain production. However, wheat varieties such as WB9518 alsoprovide a source of breeding material that may be used to develop newwheat varieties. Plant breeding techniques known in the art and used ina wheat plant breeding program include, but are not limited to,recurrent selection, mass selection, bulk selection, mass selection,backcrossing, pedigree breeding, open pollination breeding, restrictionfragment length polymorphism enhanced selection, genetic marker enhancedselection, making double haploids, and transformation. Often,combinations of these techniques are used. The development of wheatvarieties in a plant breeding program requires, in general, thedevelopment and evaluation of homozygous varieties. There are manyanalytical methods available to evaluate a new variety. The oldest andmost traditional method of analysis is the observation of phenotypictraits but genotypic analysis may also be used.

Additional Breeding Methods

In an embodiment, this invention is directed to methods for producing awheat plant by crossing a first parent wheat plant with a second parentwheat plant wherein either the first or second parent wheat plant iscultivar WB9518. The other parent may be any other wheat plant, such asa wheat plant that is part of a synthetic or natural population. Anysuch methods using wheat variety WB9518 are part of this invention:selfing, sibbing, backcrosses, mass selection, pedigree breeding, bulkselection, hybrid production, and crosses to populations. These methodsare well known in the art and some of the more commonly used breedingmethods are described below.

The following describes breeding methods that may be used with wheatcultivar WB9518 in the development of further wheat plants. One suchembodiment is a method for developing a cultivar WB9518 progeny wheatplant in a wheat plant breeding program comprising: obtaining the wheatplant, or a part thereof, of cultivar WB9518 utilizing said plant orplant part as a source of breeding material and selecting a wheatcultivar WB9518 progeny plant with molecular markers in common withcultivar WB9518 and/or with morphological and/or physiologicalcharacteristics selected from the characteristics listed in the Tablesherein. Breeding steps that may be used in the wheat plant breedingprogram include pedigree breeding, backcros sing, mutation breeding, andrecurrent selection. In conjunction with these steps, techniques such asRFLP-enhanced selection, genetic marker enhanced selection (e.g., SSRmarkers) and the making of double haploids may be utilized.

Another method involves producing a population of wheat cultivar WB9518progeny wheat plants, comprising crossing cultivar WB9518 with anotherwheat plant, thereby producing a population of wheat plants, which, onaverage, derive 50% of their alleles from wheat cultivar WB9518. A plantof this population may be selected and repeatedly selfed or sibbed witha wheat cultivar resulting from these successive filial generations. Oneembodiment of this invention is the wheat cultivar produced by thismethod and that has obtained at least 50% of its alleles from wheatcultivar WB9518.

One of ordinary skill in the art of plant breeding would know how toevaluate the traits of two plant varieties to determine if there is nosignificant difference between the two traits expressed by thosevarieties. Thus the invention includes wheat cultivar WB9518 progenywheat plants comprising a combination of at least two cultivar WB9518traits selected from the group consisting of those listed in the Tablesherein, so that said progeny wheat plant is not significantly differentfor said traits than wheat cultivar WB9518. Using techniques describedherein, molecular markers may be used to identify said progeny plant asa wheat cultivar WB9518 progeny plant. Mean trait values may be used todetermine whether trait differences are significant, and the traits maybe measured on plants grown under the same environmental conditions.Once such a variety is developed its value is substantial, as it isimportant to advance the germplasm base as a whole in order to maintainor improve traits such as yield, disease resistance or tolerance, pestresistance or tolerance, and plant performance in extreme environmentalconditions.

Progeny of wheat cultivar WB9518 may also be characterized through theirfilial relationship with wheat cultivar WB9518, as for example, beingwithin a certain number of breeding crosses of wheat cultivar WB9518. Abreeding cross is a cross made to introduce new genetics into theprogeny, and is distinguished from a cross, such as a self or a sibcross, made to select among existing genetic alleles. The lower thenumber of breeding crosses in the pedigree, the closer the relationshipbetween wheat cultivar WB9518 and its progeny. For example, progenyproduced by the methods described herein may be within 1, 2, 3, 4 or 5breeding crosses of wheat cultivar WB9518.

Pedigree Breeding

Pedigree breeding starts with the crossing of two genotypes, such asWB9518 and another wheat variety having one or more desirablecharacteristics that is lacking or which complements WB9518. If the twooriginal parents do not provide all the desired characteristics, othersources can be included in the breeding population. In the pedigreemethod, superior plants are selfed and selected in successive filialgenerations. In the succeeding filial generations the heterozygouscondition gives way to homogeneous varieties as a result ofself-pollination and selection. Typically in the pedigree method ofbreeding, five or more successive filial generations of selfing andselection is practiced: F1 to F2; F2 to F3; F3 to F4; F4 to F5, etc.After a sufficient amount of inbreeding, successive filial generationswill serve to increase seed of the developed variety. In an embodiment,the developed variety comprises homozygous alleles at about 95% or moreof its loci.

In addition to being used to create a backcross conversion, backcrossingcan also be used in combination with pedigree breeding. As discussedpreviously, backcrossing can be used to transfer one or morespecifically desirable traits from one variety, the donor parent, to adeveloped variety called the recurrent parent, which has overall goodagronomic characteristics yet lacks that desirable trait or traits.However, the same procedure can be used to move the progeny toward thegenotype of the recurrent parent but at the same time retain manycomponents of the non-recurrent parent by stopping the backcrossing atan early stage and proceeding with selling and selection. For example, awheat variety may be crossed with another variety to produce a firstgeneration progeny plant. The first generation progeny plant may then bebackcrossed to one of its parent varieties to create a BC1 or BC2.Progeny are selfed and selected so that the newly developed variety hasmany of the attributes of the recurrent parent and yet several of thedesired attributes of the non-recurrent parent. This approach leveragesthe value and strengths of the recurrent parent for use in new wheatvarieties.

Therefore, an embodiment of this invention is a method of making abackcross conversion of wheat cultivar WB9518 comprising the steps ofcrossing a plant of wheat cultivar WB9518 with a donor plant comprisinga desired trait, selecting an F1 progeny plant comprising the desiredtrait, and backcrossing the selected F1 progeny plant to a plant ofwheat cultivar WB9518. This method may further comprise the step ofobtaining a molecular marker profile of wheat cultivar WB9518 and usingthe molecular marker profile to select for a progeny plant with thedesired trait and the molecular marker profile of WB9518. In oneembodiment, the desired trait is a mutant gene or transgene present inthe donor parent.

Recurrent Selection and Mass Selection

Recurrent selection is a method used in a plant breeding program toimprove a population of plants. WB9518 is suitable for use in arecurrent selection program. The method entails individual plants crosspollinating with each other to form progeny. The progeny are grown andthe superior progeny selected by any number of selection methods, whichinclude individual plant, half-sib progeny, full-sib progeny and selfedprogeny. The selected progeny are cross pollinated with each other toform progeny for another population. This population is planted andagain superior plants are selected to cross pollinate with each other.Recurrent selection is a cyclical process and therefore can be repeatedas many times as desired. The objective of recurrent selection is toimprove the traits of a population. The improved population can then beused as a source of breeding material to obtain new varieties forcommercial or breeding use, including the production of a syntheticcultivar. A synthetic cultivar is the resultant progeny formed by theintercros sing of several selected varieties.

Mass selection is a useful technique when used in conjunction withmolecular marker enhanced selection. In mass selection, seeds fromindividuals are selected based on phenotype or genotype. These selectedseeds are then bulked and used to grow the next generation. Bulkselection requires growing a population of plants in a bulk plot,allowing the plants to self-pollinate, harvesting the seed in bulk andthen using a sample of the seed harvested in bulk to plant the nextgeneration. Also, instead of self pollination, directed pollinationcould be used as part of the breeding program.

Mutation Breeding

Mutation breeding is another method of introducing new traits into wheatcultivar WB9518. Mutations that occur spontaneously or are artificiallyinduced can be useful sources of variability for a plant breeder. Thegoal of artificial mutagenesis is to increase the rate of mutation for adesired characteristic. Mutation rates can be increased by manydifferent means including temperature, long-term seed storage, tissueculture conditions, radiation, such as X-rays, Gamma rays (e.g. cobalt60 or cesium 137), neutrons, (product of nuclear fission by uranium 235in an atomic reactor), Beta radiation (emitted from radioisotopes suchas phosphorus 32 or carbon 14), or ultraviolet radiation (optionallyfrom 2500 to 2900 nm), or chemical mutagens (such as base analogues(5-bromo-uracil), related compounds (8-ethoxy caffeine), antibiotics(streptonigrin), alkylating agents (sulfur mustards, nitrogen mustards,epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones),azide, hydroxylamine, nitrous acid, or acridines. Once a desired traitis observed through mutagenesis the trait may then be incorporated intoexisting germplasm by traditional breeding techniques. In addition,mutations created in other wheat plants may be used to produce abackcross conversion of wheat cultivar WB9518 that comprises suchmutation. Further embodiments of the invention are the treatment ofWB9518 with a mutagen and the plant produced by mutagenesis of WB9518.

Breeding with Molecular Markers

Molecular markers, which include markers identified through the use oftechniques such as Isozyme Electrophoresis, Restriction Fragment LengthPolymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats(SSRs) and Single Nucleotide Polymorphisms (SNPs), may be used in plantbreeding methods utilizing wheat cultivar WB9518. IsozymeElectrophoresis and RFLPs have been widely used to determine geneticcomposition.

SSR technology is currently the most efficient and practical markertechnology; more marker loci can be routinely used and more alleles permarker locus can be found using SSRs in comparison to RFLPs. SingleNucleotide Polymorphisms (SNPs) may also be used to identify the uniquegenetic composition of the invention and progeny varieties retainingthat unique genetic composition. Various molecular marker techniques maybe used in combination to enhance overall resolution. Wheat DNAmolecular marker linkage maps have been rapidly constructed and widelyimplemented in genetic studies.

One use of molecular markers is QTL mapping. QTL mapping is the use ofmarkers which are known to be closely linked to alleles that havemeasurable effects on a quantitative trait. Selection in the breedingprocess is based upon the accumulation of markers linked to the positiveeffecting alleles and/or the elimination of the markers linked to thenegative effecting alleles from the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against thegenome of the donor parent. Using this procedure can minimize the amountof genome from the donor parent that remains in the selected plants. Itcan also be used to reduce the number of crosses back to the recurrentparent needed in a backcrossing program. The use of molecular markers inthe selection process is often called genetic marker enhanced selection.Molecular markers may also be used to identify and exclude certainsources of germplasm as parental varieties or ancestors of a plant byproviding a means of tracking genetic profiles through crosses.

Production of Double Haploids

The production of double haploids can also be used for the developmentof plants with a homozygous phenotype in the breeding program. Forexample, a wheat plant for which wheat cultivar WB9518 is a parent canbe used to produce double haploid plants. Double haploids are producedby the doubling of a set of chromosomes (1N) from a heterozygous plantto produce a completely homozygous individual. This can be advantageousbecause the process omits the generations of selfing needed to obtain ahomozygous plant from a heterozygous source.

Haploid induction systems have been developed for various plants toproduce haploid tissues, plants and seeds. The haploid induction systemcan produce haploid plants from any genotype by crossing a selected line(as female) with an inducer line. Such inducer lines for maize includeStock 6. Methods for obtaining haploid plants have also been disclosedin the art.

Thus, an embodiment of this invention is a process for making asubstantially homozygous WB9518 progeny plant by producing or obtaininga seed from the cross of WB9518 and another wheat plant and applyingdouble haploid methods to the F1 seed or F1 plant, or to any successivefilial generation. Based on studies in maize and currently beingconducted in wheat, such methods would decrease the number ofgenerations required to produce a variety with similar genetics orcharacteristics to WB9518.

In particular, a process of making seed retaining the molecular markerprofile of wheat cultivar WB9518 is contemplated, such processcomprising obtaining or producing F1 seed for which wheat cultivarWB9518 is a parent, inducing doubled haploids to create progeny withoutthe occurrence of meiotic segregation, obtaining the molecular markerprofile of wheat cultivar WB9518, and selecting progeny that retain themolecular marker profile of WB9518. Descriptions of other breedingmethods that are commonly used for different traits and crops are known.

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of wheat andregeneration of plants therefrom is well known and widely published.Thus, another aspect of this invention is to provide cells which upongrowth and differentiation produce wheat plants having essentially allof the physiological and morphological characteristics of wheat cultivarWB9518. Means for preparing and maintaining plant tissue culture arewell known in the art. By way of example, a tissue culture comprisingorgans has been used to produce regenerated plants.

In Tables 2 and 3, yield, quality and agronomic characteristicscollected in 2011 and 2012, respectively, in the Pacific Northwest ofthe United States for wheat cultivar WB9518 are compared to twocommercial check cultivars. Column 1 shows the cultivar, column 2 showsthe yield as a percent of the trial average, column 3 shows the testweight of harvested grain in pounds per bushel, column 4 shows theJulian flowering date when 50% of the variety flowers, column 5 showsthe plant height in centimeters, column 6 shows the grain protein % on a12% moisture basis, column 7 shows the SDS Sedimentation in mm andcolumn 8 shows the stripe rust rating on a scale of 1 (least) to 9(most).

TABLE 2 Comparative Data for Cultivar WB9518 and Selected Varieties in2011 in the Pacific Northwest of the US 1 2 3 4 5 6 7 8 CharacteristicTest Flowering Plant Grain SDS Stripe Yield Weight Date Height ProteinSedimentation Rust Unit of Measure % of % Average lbs/bu Julian cm 12%mb mm 1-9* EXPRESSO 110.4 62.7 193 83.8 15.5 105 1 HANK 86.1 62.3 19088.9 14.4 107 5 Average 100.0 61.7 191.5 77.6 13.1 107 2 WB9518 111.562.9 192 81.3 15.2 104 1 No. of Tested 18 12 3 18 9 9 3 Replications*Disease rating scale: 1 = least disease and 9 = most disease.

TABLE 3 Comparative Data for Cultivar WB9518 and Selected Varieties in2012 in the Pacific Northwest of the US 1 2 3 4 5 6 7 8 CharacteristicTest Flowering Plant Grain SDS Stripe Yield Weight Date Height ProteinSedimentation Rust Unit of Measure % of % Average lbs/bu Julian cm 12%mb mm 1-9* WESTBRED 86.0 59.5 184 79.5 14.1 111.3 9 936 VOLT 104.9 63.5192 89.2 13.5 113.1 1 Average 100.0 61.9 186 83.1 14.1 108.4 2.4 WB9518107.1 62.4 188 80.8 14.1 105.2 1 No. of Tested 21 7 3 14 10 10 3Replications *Disease rating scale: 1 = least disease and 9 = mostdisease.

DEFINITIONS

In the description and tables, a number of terms are used. In order toprovide a clear and consistent understanding of the specification andclaims, the following definitions are provided:

A: When used in conjunction with the word “comprising” or other openlanguage in the claims, the words “a” and “an” denote “one or more.”

About: Refers to embodiments or values that include the standarddeviation of the mean for a given item being measured.

Allele: Any of one or more alternative forms of a gene locus, all ofwhich relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Aphids: Aphid resistance is scored on a scale from 1 to 9; a score of 4or less indicates resistance. Varieties scored as 1 to 5 appear normaland healthy, with numbers of aphids increasing from none to up to 300per plant. A score of 7 indicates that there are 301 to 800 aphids perplant and that the plants show slight signs of infestation. A score of 9indicates severe infestation and stunted plants with severely curled andyellow leaves.

Awn: Awn is intended to mean the elongated needle-like appendages on theflower- and seed-bearing head at the top of the cereal grain plant(e.g., wheat, common wheat, rye). Awns are attached to the lemmas.Lemmas enclose the stamen and the stigma as part of the florets. Floretsare grouped in spikelets, which in turn together comprise the head.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Cell: As used herein, the term cell includes a plant cell, whetherisolated, in tissue culture, or incorporated in a plant or plant part.

Chromatography: A technique wherein a mixture of dissolved substancesare bound to a solid support followed by passing a column of fluidacross the solid support and varying the composition of the fluid. Thecomponents of the mixture are separated by selective elution.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Disease Resistance: As used herein, the term disease resistance ordisease resistant is defined as the ability of plants to restrict theactivities of a specified disease, such as a fungus, virus, orbacterium.

Disease Tolerance: As used herein, the term disease tolerance or diseasetolerant is defined as the ability of plants to endure a specifieddisease (such as a fungus, virus, or bacterium) or an adverseenvironmental condition and still perform and produce in spite of thisdisorder.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Embryo: The embryo is the small plant contained within a mature seed.

Emergence (EMR): The emergence score describes the ability of a seed toemerge from the soil after planting. Each genotype is given a 1 to 9score based on its percent of emergence. A score of 1 indicates anexcellent rate and percent of emergence, an intermediate score of 5indicates an average rating and a 9 score indicates a very poor rate andpercent of emergence.

Enzymes: Molecules which can act as catalysts in biological reactions.

Essentially all of the physiological and morphological characteristics:The characteristics of a plant are recovered that are otherwise presentwhen compared in the same environment, other than occasional varianttraits that might arise during backcrossing or direct introduction of atransgene.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Gene Converted (Conversion): Gene conversion or a gene converted plantrefers to plants that are developed by backcrossing, geneticengineering, or mutation, wherein essentially all of the desiredmorphological and physiological characteristics of a variety arerecovered, in addition to the one or more traits transferred into thevariety via the back-crossing technique, genetic engineering, ormutation.

Gene Silencing: Gene silencing refers to the interruption or suppressionof the expression of a gene at the level of transcription ortranslation.

Genotype: The genetic constitution of a cell or organism.

Glume Blotch: Glume Blotch is a disease of wheat characterized by small,irregular gray to brown spots or blotches on the glumes, althoughinfections may also occur at the nodes. The disease is caused by thefungus Stagonosporum nodorum (may also be referred to as Septorianodorum). Resistance to this disease is scored on scales that reflectthe observed extent of the disease on the leaves of the plant. Ratingscales may differ but in general a low number indicates resistance andhigher number suggests different levels of susceptibility.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Head: As used herein, the term head refers to a group of spikelets atthe top of one plant stem. The term spike also refers to the head of aplant located at the top of one plant stem.

Herbicide Resistance: As used herein, the term herbicide resistance orherbicide resistant is defined as the ability of plants to survive andreproduce following exposure to a dose of herbicide that would normallybe lethal to the plant.

Herbicide Tolerance: As used herein, the term herbicide tolerance orherbicide tolerant is defined as the ability of plants to survive andreproduce after herbicide treatment.

Insect Resistance: As used herein, the term disease resistance ordisease resistant is defined as the ability of plants to restrict theactivities of a specified insect or pest.

Insect Tolerance: As used herein, the term disease tolerance or diseasetolerant is defined as the ability of plants to endure a specifiedinsect or pest and still perform and produce in spite of this disorder.

Kernel Weight: As used herein, the term kernel weight refers to theweight of individual kernels (also called seeds), often reported as theweight of one thousand kernels or “1000 Kernel Weight.”

Leaf Rust: Leaf Rust is a disease of wheat characterized by pustulesthat are circular or slightly elliptical, that usually do not coalesce,and contain masses of orange to orange-brown spores. The disease iscaused by the fungus Puccinia recondita f. sp. tritici. Infection sitesprimarily are found on the upper surfaces of leaves and leaf sheaths,and occasionally on the neck and awns. Resistance to this disease isscored on scales that reflect the observed extent of the disease on theleaves of the plant. Rating scales may differ but in general a lownumber indicates resistance and higher number suggests different levelsof susceptibility.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Locus: A locus is a position on a genomic sequence that is usually foundby a point of reference, for example, the position of a DNA sequencethat is a gene, or part of a gene or intergenic region. A locus confersone or more traits such as, for example, male sterility, herbicidetolerance or resistance, insect resistance or tolerance, diseaseresistance or tolerance, modified fatty acid metabolism, modified phyticacid metabolism, modified carbohydrate metabolism or modified proteinmetabolism. The trait may be, for example, conferred by a naturallyoccurring gene introduced into the genome of the variety bybackcrossing, a natural or induced mutation, or a transgene introducedthrough genetic transformation techniques. A locus may comprise one ormore alleles integrated at a single chromosomal location.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Maturity: As used herein, the term maturity refers to the stage of plantgrowth at which the development of the kernels is complete.

Or: As used herein is meant to mean “and/or” and be interchangeabletherewith unless explicitly indicated to refer to the alternative only.

Pedigree Distance: Pedigree distance is the relationship amonggenerations based on their ancestral links as evidenced in pedigrees. Itmay be measured by the distance of the pedigree from a given startingpoint in the ancestry.

Percent Identity: Percent identity, as used herein, refers to thecomparison of the homozygous alleles of two wheat varieties. Percentidentity is determined by comparing a statistically significant numberof the homozygous alleles of two developed varieties. For example, apercent identity of 90% between wheat variety 1 and wheat variety 2means that the two varieties have the same allele at 90% of their loci.

Percent Similarity: Percent similarity as used herein refers to thecomparison of the homozygous alleles of a wheat variety such as WB9518with another plant, and if the homozygous allele of WB9518 matches atleast one of the alleles from the other plant then they are scored assimilar. Percent similarity is determined by comparing a statisticallysignificant number of loci and recording the number of loci with similaralleles as a percentage. A percent similarity of 90% between WB9518 andanother plant means that WB9518 matches at least one of the alleles ofthe other plant at 90% of the loci.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Plant: As used herein, the term plant includes reference to an immatureor mature whole plant, including a plant from which seed, grain, oranthers have been removed. A seed or embryo that will produce the plantis also considered to be a plant.

Plant Height (Hgt): As used herein, the term plant height is defined asthe average height in inches or centimeters of a group of plants, asmeasured from the ground level to the tip of the head, excluding awns.

Plant Parts: As used herein, the term plant parts (or reference to “awheat plant, or a part thereof”) includes, but is not limited to,protoplasts, callus, leaves, stems, roots, root tips, anthers, pistils,seed, grain, pericarp, embryo, pollen, ovules, cotyledon, hypocotyl,spike, floret, awn, lemma, shoot, tissue, petiole, cells, andmeristematic cells.

Powdery Mildew: Powdery Mildew is a disease of wheat characterized bywhite to pale gray, fuzzy or powdery colonies of mycelia, and conidia onthe upper surfaces of leaves and leaf sheaths (especially on lowerleaves), and sometimes on the spikes. The disease is caused by thefungus Erysiphe graminis f. sp. tritici. Older fungal tissue isyellowish gray. This superficial fungal material can be rubbed offeasily with the fingers. Host tissue beneath the fungal material becomeschlorotic or necrotic and, with severe infections, the leaves may die.Eventually, black spherical fruiting structures may develop in themycelia, and can be seen without magnification. Resistance to thisdisease is scored on scales that reflect the observed extent of thedisease on the leaves of the plant. Rating scales may differ but ingeneral a low number indicates resistance and higher number suggestsdifferent levels of susceptibility.

Progeny: As used herein, progeny includes an F1 wheat plant producedfrom the cross of two wheat plants where at least one plant includeswheat cultivar WB9518.

Progeny further includes, but is not limited to, subsequent F₂, F₃, F₄,F₅, F₆, F₇, F₈, F₉ and F₁₀ generational crosses with the recurrentparental line.

Quantitative Trait Loci (QTL): Quantitative trait loci refer to geneticloci that control to some degree numerically representable traits thatare usually continuously distributed.

Regeneration: The development of a plant from tissue culture.

Rhizoctonia Root Rot: Rhizoctonia Root Rot is a disease of wheatcharacterized by sharp eyespot lesions that develop on basal leafsheaths. The disease is caused by the fungus Rhizoctonia solani. Thelesion margins are dark brown with pale, straw-colored centers and themycelia often present in the centers of lesions are easily removed byrubbing. Roots can also be affected, usually becoming brown in color andreduced in number. The disease can cause stunting and a reduction in thenumber of tillers. Resistance to this disease is scored on scales thatreflect the observed extent of the disease on the leaf sheaths of theplant and on reduced vigor of the plant. Rating scales may differ but ingeneral a low number indicates resistance and higher number suggestsdifferent levels of susceptibility.

Scab or Head Blight: Scab or Head Blight a disease of wheatcharacterized by florets (especially the outer glumes) that becomeslightly darkened and oily in appearance. The disease is caused by thefungus Fusarium which has numerous species. Spores are produced that cangive the spike and shriveled, infected kernels a bright pinkish color.Spores can produce a toxin, deoxynivalenol which can be measured with achemical test. Resistance to this disease can be measured in three ways:the extent of the disease on the spikes of the plant, the percentkernels which are visibly shriveled and the amount of deoxynivalenol inthe kernels. Rating scales may differ but in general a low numberindicates resistance and higher number suggests different levels ofsusceptibility.

SDS Sedimentation: SDS sedimentation (sodium dodecyl sedimentation) testvalues are a measure of the end-use mixing and handling properties ofbread dough and their relation to bread-making quality as a result ofthe dough's gluten quality. Higher SDS sedimentation levels reflecthigher gluten quality.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Septoria Leaf Blotch or Speckled Leaf Blotch: Speckled leaf blotch is adisease of wheat, common wheat and durum wheat characterized byirregularly shaped blotches that are at first yellow and then turnreddish brown with grayish brown dry centers, caused by the rust fungusSeptoria tritici. Resistance to this disease is scored on scales thatreflect the observed extent of the disease on the leaves of the plant.Rating scales may differ but in general a low number indicatesresistance and higher number suggests different levels ofsusceptibility.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing and/or by genetictransformation to introduce a given locus that is transgenic in origin,wherein essentially all of the morphological and physiologicalcharacteristics of a wheat cultivar are recovered in addition to thecharacteristics of the locus transferred into the variety via thebackcrossing technique or by genetic transformation. It is understoodthat once introduced into any wheat plant genome, a locus that istransgenic in origin (transgene), can be introduced by backcrossing aswith any other locus.

Soil Born Mosaic Virus: Soil born mosaic virus is a disease of wheatcharacterized by mild green to yellow mosaic, yellow-green mottling,dashes, and parallel streaks, most clearly visible on the youngest leaf.Reddish streaking and necrosis at leaf tips sometimes occurs. Stuntingcan be moderate to severe, depending on the variety. The disease iscaused by a virus which is transmitted by a soilborne fungus-likeorganism, Polymyxa graminis, which makes swimming spores that infect theroots of wheat. Resistance to this disease is scored on scales thatreflect the observed extent of the disease on the young plants. Ratingscales may differ, but in general, a low number indicates resistance anda higher number suggests different levels of susceptibility.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Stem Rust: Stem Rust is a disease of wheat characterized by pustulescontaining masses of spores that are dark reddish brown, and may occuron both sides of the leaves, on the stems, and on the spikes. Thedisease is caused by the fungus Puccinia graminis f. sp. Tritici.Resistance to this disease is scored on scales that reflect the observedextent of the disease on the leaves of the plant. Rating scales maydiffer, but in general, a low number indicates resistance and a highernumber suggests different levels of susceptibility.

Stripe Rust: Stripe rust is a disease of wheat, common wheat, durumwheat, and barley characterized by elongated rows of yellow spores onthe affected parts, caused by a rust fungus, Puccinia striiformis.Resistance to this disease is scored on scales that reflect the observedextent of the disease on the leaves of the plant. Rating scales maydiffer, but in general, a low number indicates resistance and a highernumber suggests different levels of susceptibility.

Test Weight (TWT): As used herein, the term test weight is a measure ofdensity that refers to the weight in pounds of the amount of kernelscontained in a bushel unit of volume.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant. Exemplary types of tissue cultures are protoplasts, calli, plantclumps, and plant cells that can generate tissue culture that are intactin plants or parts of plants, such as embryos, pollen, ovules, pericarp,flowers, florets, heads, spikelets, seeds, leaves, stems, roots, roottips, anthers, pistils, awns, stems, and the like.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a wheat plant by transformation.

DEPOSIT INFORMATION

A deposit of the wheat cultivar WB9518, which is disclosed herein aboveand referenced in the claims, will be made with the American TypeCulture Collection (ATCC), 10801 University Blvd., Manassas, Va.20110-2209. The date of deposit is ______ and the accession number forthose deposited seeds of wheat cultivar WB9518 is ATCC Accession No.______. All restrictions upon the deposit have been removed, and thedeposit is intended to meet all of the requirements of 37 C.F.R.§1.801-1.809. The deposit will be maintained in the depository for aperiod of 30 years, or 5 years after the last request, or for theeffective life of the patent, whichever is longer, and will be replacedif necessary during that period.

All of the compositions and methods disclosed and claimed herein can bemade and executed without undue experimentation in light of the presentdisclosure. While the compositions and methods of this invention havebeen described in terms of the foregoing illustrative embodiments, itwill be apparent to those of skill in the art that variations, changes,modifications, and alterations may be applied to the composition,methods, and in the steps or in the sequence of steps of the methodsdescribed herein, without departing from the true concept, spirit, andscope of the invention. More specifically, it will be apparent thatcertain agents that are both chemically and physiologically related maybe substituted for the agents described herein while the same or similarresults would be achieved. All such similar substitutes andmodifications apparent to those skilled in the art are deemed to bewithin the spirit, scope, and concept of the invention as defined by theappended claims.

The references cited herein, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference.

1. A plant of wheat cultivar WB9518, wherein a sample of seed of saidvariety has been deposited under ATCC Accession No. ______.
 2. A plantpart of the plant of claim 1, wherein the plant part comprises at leastone cell of said plant.
 3. The plant part of claim 2, further defined ashead, awn, leaf, pollen, ovule, embryo, cotyledon, hypocotyl,meristematic cell, root, root tip, pistil, anther, floret, seed,pericarp, spike, stem, and callus.
 4. A tissue culture produced from theplant part of claim
 3. 5. A wheat plant regenerated from the tissueculture of claim
 4. 6. A seed of wheat cultivar WB9518, wherein a sampleof seed of said variety has been deposited under ATCC Accession No.______.
 7. A method of producing wheat seed, wherein the methodcomprises crossing the plant of claim 1 with itself or a second wheatplant.
 8. The method of claim 7, wherein the method is further definedas comprising crossing the plant of wheat cultivar WB9518 with a second,distinct wheat plant to produce an F₁ hybrid wheat seed.
 9. An F₁ hybridwheat seed produced by the method of claim
 8. 10. An F₁ hybrid wheatplant produced by growing the seed of claim
 9. 11. A compositioncomprising the seed of claim 6 comprised in plant seed growth media,wherein a sample of seed of said variety has been deposited under ATCCAccession No. ______.
 12. The composition of claim 11, wherein thegrowth media is soil or a synthetic cultivation medium.
 13. A plantproduced by introducing a single locus conversion into wheat cultivarWB9518, or a selfed progeny thereof comprising the single locusconversion, wherein the single locus conversion was introduced intowheat cultivar WB9518 by backcrossing or genetic transformation andwherein a sample of seed of wheat cultivar WB9518 has been depositedunder ATCC Accession No. ______.
 14. The plant of claim 13, wherein thesingle locus conversion comprises a transgene.
 15. A seed that producesthe plant of claim 13
 16. The seed of claim 15, wherein the single locusconfers a trait selected from the group consisting of male sterility,herbicide tolerance, insect resistance, pest resistance, diseaseresistance, modified fatty acid metabolism, abiotic stress resistance,altered seed amino acid composition, site-specific geneticrecombination, and modified carbohydrate metabolism.
 17. The seed ofclaim 16, wherein the single locus confers tolerance to an herbicideselected from the group consisting of glyphosate, sulfonylurea,imidazolinone, dicamba, glufosinate, phenoxy proprionic acid,L-phosphinothricin, cyclohexone, cyclohexanedione, triazine, andbenzonitrile.
 18. The seed of claim 16, wherein the single locus furthercomprises a gene encoding a Bacillus thuringiensis (Bt) endotoxin. 19.The seed of claim 16, wherein the single locus further comprises a geneencoding a protein selected from the group consisting of glutenins,gliadins, phytase, fructosyltransferase, levansucrase, α-amylase,invertase and starch branching enzyme or encoding an antisense ofstearyl-ACP desaturase.
 20. The seed of claim 15, wherein the singlelocus conversion comprises a transgene.
 21. The method of claim 8,wherein the method further comprises: (a) crossing a plant grown fromsaid F₁ hybrid wheat seed with itself or a different wheat plant toproduce a seed of a progeny plant of a subsequent generation; (b)growing a progeny plant of a subsequent generation from said seed of aprogeny plant of a subsequent generation and crossing the progeny plantof a subsequent generation with itself or a second plant to produce aprogeny plant of a further subsequent generation; and (c) repeatingsteps (a) and (b) using said progeny plant of a further subsequentgeneration from step (b) in place of the plant grown from said F₁ hybridwheat seed in step (a), wherein steps (a) and (b) are repeated withsufficient inbreeding to produce an inbred wheat plant derived from thewheat cultivar WB9518.
 22. A method of producing a commodity plantproduct comprising collecting the commodity plant product from the plantof claim
 1. 23. The method of claim 22, wherein the commodity plantproduct is grain, flour, baked goods, cereals, pasta, beverages,livestock feed, biofuel, straw, construction materials, and starches.24. A wheat commodity plant product produced by the method of claim 23,wherein the commodity plant product comprises at least one cell of wheatcultivar WB9518.